RETNO SINTOWATI

Purin, Pirimidin, Asam nukleat

I. Pendahuluan
II. Basa Purin dan Basa Pirimidin
III. Nukleosida
IV. Nukleotida
V. Asam Nukleat
 Kepentingan biomedis

 kemampuan nukleotid menyerap sinar UV 

menjadikan sinar UV  sbg unsur mutagen yg
potent
 senyawa analog purin & pirimidin disintesis
secara kimiawi  digunakan untuk terapi
penyakit kanker
 Sel yg akan membelah, selnya mengalami
replikasi
 Katabolisme purin  xantin  asam urat
 I. Pendahuluan
 Nukleoprotein
 protein majemuk
 terdapat dalam inti sel
 penyusun material inti sel
 tersusun atas :
protein: histon / protamin
gugus non protein
 semua sel hidup mengandung nukleoprotein
 Kromatin
 Konstituen penting dalam sel, tersusun atas
nukleoprotein
 kelainan pembentukan nukleoprotein
perubahan dlm pertumbuhan /reproduksi sel
sebab kromatin pembelahan sel
 Nukleoprotein
Enzim proteolitik
- Protein sederhana - Asam nukleat
trtm: histon & protamin (RNase / DNase)

polinukleotida
polinukleotidase
(fosfoesterse)
mononukleotida
purin/pirimidin nukleotidase (fosfatase)

nukleosida asam fosfat

(fosforilase)
basa purin/basa pirimidin gula (ribosa/deoksi ribosa)
 Nukleosida Purin

Adenosin Guanosin
adenosin deaminase purin nukleosida fosforilase
NH3 Inosin Guanin
purin nukleosida
fosforilase
Hipoxantin
Xantin oksidase
Xantin + O2 asam urat urin
Xantin oksidase
 Penyelamatan Purin
Untuk pembentukan nukleotida bbrp jaringan manusia
tgt pd purin /purin ribonukleosida eksogen
Ada 2 jalan penyelamatan :
1. Fosforibosilasi purin bebas dgn enzim yang
membutuhkan PRPP sbg donor ribosa fosfat
2. Fosforilasi nukleosida purin pd gugus 5’-hidroksi oleh
enzim adenosin kinase
Contoh :
1. Adenin menjadi AMP dg enzim adenin fosforibosa
transferase
2. Hipoxantin dan Guanin menjadi IMP dan GMP dengan
enzim hipoxantin-guanin fosforibosa transferase
 Nukleosida pirimidin
Sitidin Deoksisitidin
deaminasi

Uridin deaminasi
Deoksiuridin Deoksitimidin
fosforilse
Urasil dan Timin
degradasi

-alanin + NH4+ +CO2 degradasi

As. -amino isobutirat + NH4+ +CO2

urine
Banyak tdp pd penderita tumor yang dikhemoterapi
( DNA degradasi )
 II. Basa Purin Dan Basa Pirimidin
Basa Purin
 Basa heterosiklis , molekul planar , aromatik
 sebag. besar tdp. dlm sel dlm. bentuk asam nukleat
 disintesis secara de novo, dg sumber :
 N1 dari as. Aspartat,
 C6 dr CO2 respirasi,
 N7 dari glisin,
 C8 dari N5,N10-metenilFH4,
 N3 dan N9 dari nitrogen amida der. Glutamin dan
 C2 dari N-formil -tetrahidroksi folat
 Diberi nomor mulai atom N1 kearah kebalikan jarum jam
 Basa Pirimidin
 nama lebih panjang dari purin ttp hy mgd 6
atom yang heterosiklis, sedang purin
nama pendek mgd 9 atom heterosiklis

 Sitosin (S): tdp dlm semua asam nukleat, kecuali DNA

virus ttt
 Timin (T) : trtm tdp dlm as nukleat
yg mgd gula deoksiribosa (DNA);
sedikit tdp dlm tRNA
 Urasil (U): tdp dalam RNA
Nitrogenous Bases
 Planar, aromatic, and heterocyclic
 Derived from purine or pyrimidine
 Numbering of bases is “unprimed”
Nucleic Acid Bases
Purines Pyrimidines
Sugars
 Pentoses (5-C sugars)
 Numbering of sugars is “primed”
Sugars
D-Ribose and 2’-Deoxyribose

*Lacks a 2’-OH group

Nucleosides
 Result from linking one of the sugars with a purine or
pyrimidine base through an N-glycosidic linkage

 Purines bond to the C1’ carbon of the sugar at their N9

atoms
 Pyrimidines bond to the C1’ carbon of the sugar at their
N1 atoms
Nucleosides
Phosphate Groups
 Mono-, di- or triphosphates

 Phosphates can be bonded to either C3 or C5 atoms of

the sugar
Nucleotides
 Result from linking one or more phosphates with
a nucleoside onto the 5’ end of the molecule
through esterification
 III. Nukleosida dan nukleotida
Nukleosida:
- tdr dari basa + d-ribosa  gula ribosa terikat
pada N9 basa purin dan pada N1 basa pirimidin
(kecuali U pd C5 ikatan C-C) dan semuanya
mrpk ikatan -N-glikosidik
- Nukleosida yang dimanfaatkan untuk
menghambat pertumbuhan sel:
- 5-fluorourasil - 5’-iodo-2’-deoksiuridin
- 6-tioguanin - 6-merkaptopurin
- 6-azauridin - arabinosil sitosin
Nukleotida
 Mrpk unit struktural as nukleat, mrpk komponen seluler
yg mudah dpt diketahui krn menyerap kuat sinar UV, der. basa
purin absorbsinya lebih kuat dari der. basa pirimidin
 Tersusun: - basa purin/ basa pirimidin
- gula (ribosa atau 2-deoksiribosa),
- asam fosfat
 Pengikatan fosfat: RNA pd at C3’ dari gulanya
DNA pd at C5’ dari gulanya
Basa Nukleosida Nukleotida Singkatan
RNA DNA
Adenin Adenosin Asam adenilat A dA
Guanin Guanosin Asam guanilat G dG
Sitosin Sitidin Asam Sitidilat C dC
Urasil Uridin Asam uridilat U dU
Timin Timidin Asam timidilat T dT
 Lanjutan nukleotida
- bentuk nukleotida lebih mudah larut dalam air dp nukleosida dan
basa bebasnya

Fungsi nukleotida
1. Berperan dlm metabolisme energi
Contoh: ATP - bentuk energi kimia yang diperlukan
untuk sel, dihasilkan dari fosforilasi
oksidatif atau fosforilasi tingkt substrat
- terlibat dalam kontraksi otot
- transport aktif
- mempertahankan gradien/ion
- membantu sbg donor fosfat untuk sin-
tesis nukleosida 5’-trifosfat
substrat untuk reaksi yg dikatalisis
oleh RNA dan DNA polimerase
Lanjutan fungsi nukleotida

2. Monomer unit asam nukleat: DNA dan RNA

3. Mediator fisiologik: nukleosida dan nukleotida
membantu sbg kunci mediator fisiologik proses
metabolik:
- adenosin penting dlm kontrol aliran darah koroner
- ADP  critical dlm agregasi platelet dan koagulasi
darah
- cAMP & cGMP: bekerja sbg second messenger
- GTP: diperlukan untuk capping mRNA
4. Precursor function: GTP adalah prekursor pemben-
tukan kofaktor, tetrahidrobiopterin, untuk reaksi
hidroksilasi dan generasi oksida nitrat
5. Komponen koenzim: NAD, NADP, FAD
 Lanjutan fungsi nukleotida
6. Activated intermediate
- nukleotida membantu sbg kunci activated
intermediate, diperlukan untuk berbagai reaksi
- UDP-glukosa-kinase intermidiate sintesis glikogen,
glikoprotein dll.
7. Allosteric effectors pd metabolisme nukleotida

Sintesis nukleotida
- Di dalam sel mamalia disintesis secara de novo
- Tidak semua sel mampu mensintesis nukleotid purin
Contoh; sel-sel darah merah
- Pada akhir reaksi menghasilkan ITP  prekursor
adenosin 5’-monofosfat (AMP) dan guanosin 5’-mono-
- Fosfat (GMP)
Lanjutan sintesis nukleotida
- IMP  AMP energinya dari GTP
- IMP  GMP energinya dari ATP
- Apabila terjadi defek dlm reaksi metabolik 
menyebabkan kehilangan pengaturan sintesis
nukleotida purin  over produksi purin 
asam urat meningkat
 Artritis gout
Pembentukan deoksi ribonukleotida
- pada keadaan sel tidak proliferasi  rendah
- pada saat replikasi sintesis DNA sintesis
deoksiribonukleotida meningkat untuk
mensuport sintesis DNA
 IV. Asam nukleat (Polinukleotid)
 dibentuk dari ikatan fosfodiester yg terikat
pada C3’- dan C5’ monomer yg berdekatan
 ada mol. yg berakhir 5’ dan 3’
 masing-masing mol. memp. struktur
primernya, contoh 5’-3’p Gp Gp Ap Tp Cp A
 Ada 2 macam asam nukleat besar: DNA & RNA
Nucleotide Metabolism
 PURINE RIBONUCLEOTIDES: formed de novo
 i.e., purines are not initially synthesized as free bases
 First purine derivative formed is Inosine Mono-
phosphate (IMP)
 The purine base is hypoxanthine
 AMP and GMP are formed from IMP
Purine Nucleotides
 Get broken down into Uric Acid (a purine)
Buchanan (mid 1900s) showed where purine ring
components came from:

N1: Aspartate Amine

C2, C8: Formate
N3, N9: Glutamine
C4, C5, N7: Glycine
C6: Bicarbonate Ion
 CO2
Glycine
Aspartate H
C H
N
HN C Formyl-THF
C
Formyl-THF HC C
N N
H H
Glutamine Glutamine
 H
C H
H
N Glycine C
HN C Glutamine
CH Glutamine (2) HN CH
Aspartate
HC C Aspartate
HC CH CO2
N N N10-Formyl-THF (2)
H H CO2 N
H

Purine Pyrimidine

(C5H7N4) (C4H6N2)
 O
Purine Nucleotide Synthesis OOC
COO
C
2-
O3P O CH2 H N HC N N
O C4 Aspartate ADP C4
H H  CH + ATP + Pi H
CH
5 CH2 5
H C C
OH N N
OH OH H2N SAICAR Synthetase COO H2N
-D-Ribose-5-Phosphate (R5P)
Ribose-5-Phosphate Ribose-5-Phosphate
ATP Carboxyamidoimidazole Ribotide (CAIR) 5-Aminoimidazole-4-(N-succinylocarboxamide)
Ribose
Phosphate ribotide (SAICAR)
ADP + Pi
Pyrophosphokinase AIR
Car boxylase
AMP Fumarate Adenylosuccinate
ATP Lyase
+HCO3 O
N
2-
O3P O CH2 H HC 4 C
O  O H2N N
O CH
H H C4
5
H O P O P O C CH
OH N C
5
OH
H2N N
O O H2N
Ribose-5-Phosphate
Ribose-5-Phosphate
5-Aminoimidazole Ribotide (AIR)
5-Phosphoribosyl--pyrophosphate (PRPP) 5-Aminoimidazole-4-carboxamide
ADP + Pi ribotide (AICAR)
AIR
Glutamine Synthetase N10-Formyl-
+ H2O Amidophosphoribosyl THF
ATP AICAR
Transferase
Transformylase
Glutamate H O THF
+ PPi N
H2C CH C
H2N N
2- NH2 C4
O3P O CH2
O CH
H H  C O 5
HN NH C
N
H H O C NH
OH OH H
Ribose-5-Phosphate Ribose-5-Phosphate
-5-Phosphoribosylamine (PRA)
Formylglycinamidine ribotide (FGAM) 5-Formaminoimidazole-4-carboxamide
Glycine ADP + ribotide (FAICAR)
+ ATP Glutamate + Pi
FGAM
GAR Synthetase H2O
Synthetase IMP
ATP + Cyclohydrolase
ADP Glutamine +
+ Pi H2O O
H
H2C NH2 N C N
H 2C CH HN C
4
O C CH
HC C5
2-
C O N
O3P O CH2 NH N
O N10-Formyl-THF THF O NH 2-
O3P O CH2
H H O
H H
H H H
OH
GAR Transformylase Ribose-5-Phosphate H
OH OH OH

Glycinamide Ribotide (GAR) Formylglycinamide ribotide (FGAR) Inosine Monophosphate (IMP)

IMP is converted to AMP or GMP

O GTP GDP + Pi
NH2
Aspartate Fumarate
C N C
HN C N
N C
CH CH
HC C HC C
N N N
N

Ribose-P Ribose-P
H2O + NAD+
IMP AMP
NADH + H+

O
O ATP ADP + Pi
Glutamine Glutamate C N
C N HN C
HN C CH
CH C C
C C H2N N N
HO N N

Ribose-P
Ribose-P

XMP GMP
Regulatory Control of Purine Nucleotide
Biosynthesis
 GTP is involved in AMP synthesis and ATP is involved in
GMP synthesis (reciprocal control of production)
 PRPP is a biosynthetically “central” molecule (why?)
 ADP/GDP levels – negative feedback on Ribose Phosphate
Pyrophosphokinase
 Amidophosphoribosyl transferase is activated by PRPP levels
 APRT activity has negative feedback at two sites
 ATP, ADP, AMP bound at one site
 GTP,GDP AND GMP bound at the other site
 Rate of AMP production increases with increasing
concentrations of GTP; rate of GMP production increases
with increasing concentrations of ATP
Regulatory Control of Purine Biosynthesis
 At level of IMP production:
 Independent control
 Synergistic control
 Feedforward activation by PRPP
 Below level of IMP production
 Reciprocal control

 Total amounts of purine nucleotides controlled

 Relative amounts of ATP, GTP controlled
Purine Catabolism and Salvage
 All purine degradation leads to uric acid (but it might not
stop there)
 Ingested nucleic acids are degraded by pancreatic
nucleases, and intestinal phosphodiesterases in the
intestine
 Group-specific nucleotidases and non-specific
phosphatases degrade nucleotides into nucleosides
 Direct absorption of nucleosides
 Further degradation
Nucleoside + H2O  base + ribose (nucleosidase)
Nucleoside + Pi  base + r-1-phosphate (n. phosphorylase)

NOTE: MOST INGESTED NUCLEIC ACIDS ARE DEGRADED AND

EXCRETED.
Intracellular Purine Catabolism
 Nucleotides broken into nucleosides by action of
5’-nucleotidase (hydrolysis reactions)
 Purine nucleoside phosphorylase (PNP)
 Inosine  Hypoxanthine
 Xanthosine  Xanthine
 Guanosine  Guanine
 Ribose-1-phosphate splits off
 Can be isomerized to ribose-5-phosphate

 Adenosine is deaminated to Inosine (ADA :

Adeosine deaminase)
Intracellular Purine Catabolism
 Xanthine is the point of convergence for the
metabolism of the purine bases

 Xanthine  Uric acid

 Xanthine oxidase catalyzes two reactions

 Purine ribonucleotide degradation pathway is same for

purine deoxyribonucleotides
Adenosine Degradation
Xanthosine Degradation

• Ribose sugar gets recycled (Ribose-1-Phosphate  R-5-P )

– can be incorporated into PRPP (efficiency)
• Hypoxanthine is converted to Xanthine by Xanthine Oxidase
• Guanine is converted to Xanthine by Guanine Deaminase
• Xanthine gets converted to Uric Acid by Xanthine Oxidase
 GMP AMP
↓ ↓
 Guanosin IMP
↓ ↓
↓ Inosin
↓
 Guanin Hipoxantin
↓ ↓‫٭‬

Xantin
↓‫٭‬

 Asam urat  URIN

‫ ٭‬tmp kerja xantin oxidase, dihambat oleh Alopurinol
Uric Acid Excretion
 Humans – excreted into urine as insoluble crystals
 Birds, terrestrial reptiles, some insects – excrete
insoluble crystals in paste form (hewan urikotelik)
 Excess amino N converted to uric acid
 Others – further modification :

Uric Acid  Allantoin  Allantoic Acid  Urea  Ammonia

 Ekskresi asam urat total man  400-600mg/24jam
 Aspirin dosis tinggi hambat ekskresi & reabsorpsi urat.
Purine Salvage
(Jalur penyelamatan purin)
 Adenine phosphoribosyl transferase (APRT)
Adenine + PRPP  AMP + PPi
 Hypoxanthine-Guanine phosphoribosyl transferase
(HGPRT)
Hypoxanthine + PRPP  IMP + PPi
Guanine + PRPP  GMP + PPi

(NOTE: THESE ARE ALL REVERSIBLE REACTIONS)

AMP,IMP,GMP do not need to be resynthesized de

novo !
 Di hati : nukleotida  defosforilasi nukleosida
 Sering nukleosida diputus  basa bebas + gula
 nukleosida & basa dibawa ke jar lain  refosforilasi mjd
nukleotida
 Basa bebas diselamatkan dg mereaksikan dg PRPP 
nukleotida baru
 Enzim penghemat basa tsb HGPRT
 Reaksi analog juga terjadi utk pirimidin
A CASE STUDY : GOUT
 A 45 YEAR OLD MAN AWOKE FROM SLEEP WITH A PAINFUL AND
SWOLLEN RIGHT GREAT TOE. ON THE PREVIOUS NIGHT HE HAD
EATEN A MEAL OF FRIED LIVER AND ONIONS, AFTER WHICH HE
MET WITH HIS POKER GROUP AND DRANK A NUMBER OF
BEERS.
 HE SAW HIS DOCTOR THAT MORNING, “GOUTY ARTHRITIS” WAS
DIAGNOSED, AND SOME TESTS WERE ORDERED. HIS SERUM
URIC ACID LEVEL WAS ELEVATED AT 8.0 mg/dL (NL < 7.0 mg/dL).
 THE MAN RECALLED THAT HIS FATHER AND HIS GRANDFATHER,
BOTH OF WHOM WERE ALCOHOLICS, OFTEN COMPLAINED OF
JOINT PAIN AND SWELLING IN THEIR FEET.
A CASE STUDY : GOUT
 THE DOCTOR RECOMMENDED THAT THE MAN USE
NSAIDS FOR PAIN AND SWELLING, INCREASE HIS
FLUID INTAKE (BUT NOT WITH ALCOHOL) AND REST
AND ELEVATE HIS FOOT. HE ALSO PRESCRIBED
ALLOPURINOL.
 A FEW DAYS LATER THE CONDITION HAD RESOLVED
AND ALLOPURINOL HAD BEEN STOPPED. A REPEAT
URIC ACID LEVEL WAS OBTAINED (7.1 mg/dL). THE
DOCTOR GAVE THE MAN SOME ADVICE REGARDING
LIFE STYLE CHANGES.
Gout  Impaired excretion or overproduction of uric
acid
 Uric acid crystals precipitate into joints
(Gouty Arthritis), kidneys, ureters (stones)
 Lead impairs uric acid excretion – lead
poisoning from pewter drinking goblets
 Fall of Roman Empire?
 Xanthine oxidase inhibitors inhibit
production of uric acid, and treat gout
 Allopurinol treatment – hypoxanthine analog
that binds to Xanthine Oxidase to decrease
uric acid production
ALLOPURINOL IS A XANTHINE OXIDASE INHIBITOR

A SUBSTRATE ANALOG IS CONVERTED TO AN INHIBITOR, IN THIS

CASE A “SUICIDE-INHIBITOR”
Pyrimidine Ribonucleotide Synthesis
 Uridine Monophosphate (UMP) is synthesized first
 CTP is synthesized from UMP
 Pyrimidine ring synthesis completed first; then
attached to ribose-5-phosphate

N1, C4, C5, C6 : Aspartate

C2 : HCO3-
N3 : Glutamine amide Nitrogen
 Pyrimidine Synthesis
O

2 ATP + HCO3- + Glutamine + H2O C

HN CH
2 ADP +
Carbamoyl O
Glutamate +
Phosphate C C
Pi
Synthetase II
C O N
HN CH PRPP PPi COO
2-
O3P O CH2
O
NH2
C C Orotate Phosphoribosyl H H 
N Transferase
O C O H H
H COO OH OH
O PO3-2 Orotidine-5'-monophosphate
Orotate
(OMP)
Carbamoyl Phosphate
Reduced
Aspartate Quinone OMP
Aspartate Dihydroorotate
Transcarbamoylase Decarboxylase
Dehydrogenase CO2
(ATCase)
Pi Quinone O

O C
O HN CH

C
HO C C CH
HN CH2 N
CH2 O
NH2 H2O
2-
C CH O3P O CH2
O
C CH O N H H 
Dihydroorotase
O N H COO H H
H COO OH
OH
Dihydroorotate
Carbamoyl Aspartate Uridine Monophosphate
(UMP)
UMP  UTP and CTP
 Nucleoside monophosphate kinase catalyzes
transfer of Pi to UMP to form UDP; nucleoside
diphosphate kinase catalyzes transfer of Pi from
ATP to UDP to form UTP

 CTP formed from UTP via CTP Synthetase

driven by ATP hydrolysis
 Glutamine provides amide nitrogen for C4 in animals
Regulatory Control of Pyrimidine
Synthesis
 Animals – regulation at carbamoyl phosphate
synthetase II
 UDP and UTP inhibit enzyme; ATP and PRPP activate it
 UMP and CMP competitively inhibit OMP Decarboxylase
*Purine synthesis inhibited by ADP and GDP at ribose
phosphate pyrophosphokinase step, controlling level
of PRPP  also regulates pyrimidines
Orotic Aciduria
 Caused by defect in protein chain with enzyme
activities of last two steps of pyrimidine
synthesis
 Increased excretion of orotic acid in urine
 Symptoms: retarded growth; severe anemia
 Only known inherited defect in this pathway
(all others would be lethal to fetus)
 Treat with uridine/cytidine
 HOW DOES URIDINE AND CYTIDINE
ADMINISTRATION WORK TO TREAT OROTIC
ACIDURIA?
Degradation of Pyrimidines
 CMP and UMP degraded to bases similarly to purines
 Dephosphorylation
 Deamination
 Glycosidic bond cleavage
 Uracil reduced in liver, forming -alanine
 Converted to malonyl-CoA  fatty acid synthesis for
energy metabolism
Deoxyribonucleotide Formation
 Purine/Pyrimidine degradation are the same for
ribonucleotides and deoxyribonucleotides
 Biosynthetic pathways are only for ribonucleotide
production
 Deoxyribonucleotides are synthesized from
corresponding ribonucleotides
 Pd tingkat difosfat,gugus ribosa direduksi mjd
deoksiribosa o/ enz Ribonukleotida reduktase.
ex. CTP defosforilasi  CDP reduksi  dCDP
dCTP defosforilasi dan deaminasi  dUMP
 Katabolisme pirimidin menghasilkan metabolit yg dpt
larut dalam air, yaitu : CO2, NH3, B-alanin,
B-aminoisobutirat
 Produk katab purin tidak begitu mudah larut,
kelarutan ditingkatkan dg alkalinisasi.
Daftar Bahan Makanan
 Kandungan purin tinggi ?
 Purin sedang ?
 Purin rendah ?

 Bagaimana meningkatkan kelarutan asam urat dalam

urin sehingga meningkat pula ekskresinya?
 Bagaimana cara alkalinisasi urine ?